Development of thermostabilized “ready-to-use” multiplex PCR assay for the rapid detection and distinction between Mycobacterium tuberculosis complex members and non-tuberculous mycobacteria

Tuberculosis (TB) is a worldwide health problem with significant morbidity and mortality. Current diagnosis of mycobacterial infections is largely based on clinical features, microscopy and culture. However, these methods can be time consuming, lack sensitivity and specificity. Therefore, rapid detection of both M. tuberculosis and NTM are highly desirable. Multiplex PCR assay, which is able to differentiate M. tuberculosis and NTM in clinical samples was developed in this study. It is based on the detection of both genus (targeting genes encoding 65-kD heat shock protein and dnaJ protein which are highly conserved in mycobacteria species) and species (targeting IS6110 and B9 genes which are only present in M. tuberculosis complex). The sensitivity and specificity of the multiplex PCR assay were 97.7 % and 100 %, respectively. This multiplex PCR assay has the potential to be used as a rapid diagnostic tool to detect and differentiate M. tuberculosis and NTM.